High Production of Ergothioneine in Escherichia coli using the Sulfoxide Synthase from Methylobacterium strains
High Production of Ergothioneine in Escherichia coli using the Sulfoxide Synthase from Methylobacterium strains
复制标题
DOI：
10.1021/acs.jafc.0c01846
复制
发表时间：
2020-06-10
影响因子：
6.1
通讯作者：
Dairi, Tohru
中科院分区：
文献类型：
--
作者：
Kamide, Tomoyuki;Takusagawa, Shun;Dairi, Tohru
我们以前构建了一个异源生产系统麦角硫因（ERG）在大肠杆菌中使用5个ERG生物合成基因（egtABCDE）从耻垢分枝杆菌。然而，作为ERG的ERG的中间体，大量的组氨酸甜菜碱（HER）被积累，这表明EgtB催化γ-谷氨酰半胱氨酸（γ GC）与HER的连接以产生组氨酸甜菜碱-γ-谷氨酰半胱氨酸亚砜的反应是瓶颈。在这项研究中，我们搜索了其他EgtB，并在不同的微生物中发现了许多egtB直系同源物。其中，甲基杆菌属菌株具有EgtB，其以与酸细菌CthEgtB和真菌Egt 1相似的方式催化HER直接转化为以L-半胱氨酸（L-Cys）作为硫供体的hercynylcysteine亚砜。用来自臂状甲基杆菌和拟sasicola甲基杆菌的重组EgtB进行的体外研究清楚地表明，两种酶都接受L-Cys，但不接受γ GC。我们在E. coli中的egtB。ERG生产力达到657 mg L-1。
We previously constructed a heterologous production system for ergothioneine (ERG) in Escherichia coli using five ERG biosynthesis genes (egtABCDE) from Mycobacterium smegmatis. However, significant amounts of hercynine (HER), an intermediate of ERG, as ERG were accumulated, suggesting that the reaction of EgtB catalyzing the attachment of gamma-glutamylcysteine (gamma GC) to HER to yield hercynyl-gamma-glutamylcysteine sulfoxide was a bottleneck. In this study, we searched for other EgtBs and found many egtB orthologs in diverse microorganisms. Among these, Methylobacterium strains possessed EgtBs that catalyze the direct conversion of HER into hercynylcysteine sulfoxide with L-cysteine (L-Cys) as a sulfur donor, in a manner similar to those of acidobacterial CthEgtB and fungal Egt1. An in vitro study with recombinant EgtBs from Methylobacterium brachiatum and Methylobacterium pseudosasicola clearly showed that both enzymes accepted L-Cys but not gamma GC. We reconstituted the ERG production system in E. coli with egtB from M. pseudosasicola; ERG productivity reached 657 mg L-1.